عنوان

Long-term in vivo reconstitution of hematopoiesis by human umbilical cord blood-derived highly immature stem cells

شماره

TLpq304364303

زبان متن نوشتاري يا گفتاري و مانند آن

انگلیسی

عنوان اصلي

Long-term in vivo reconstitution of hematopoiesis by human umbilical cord blood-derived highly immature stem cells

نام عام مواد

[Thesis]

نام نخستين پديدآور

N.-U.-I. Hashmi

نام ساير پديدآوران

R. E. Morris

نام ناشر، پخش کننده و غيره

University of Cincinnati

تاریخ نشرو بخش و غیره

1997

نام خاص و کميت اثر

227

جزئيات پايان نامه و نوع درجه آن

Ph.D.

کسي که مدرک را اعطا کرده

University of Cincinnati

امتياز متن

1997

متن يادداشت

Three Hematopoietic Stem Cell (HSCs) enriched fractions (i.e. CD34, CD34Lin and CD34LinRh were isolated using immunoaffinity columns and/or Fluorescence Activated Cell Sorter (FACS). The stem cell potential of the isolated cell fractions was substantiated by the short term Methylcellulose in vitro assay. The fractions were tail vein injected into sublethally irradiated Severe Combined Immune-Deficient (SCID) mice and the major hematopoietic organs of the mice were evaluated at different time points for engraftment and multilineage differentiation. High levels of multilineage engraftment were obtained as assessed by different techniques of cell and molecular biology including polymerase chain reaction (PCR) for human beta-globin gene, in situ hybridization for human specific Alu-1 sequences, human specific CD45 flow cytometry and isoelectric focusing (IEF) gel electrophoresis. No exogenous human cytokines were given to the SCID mice suggesting that the Human Umbilical Cord Blood (HUCB) derived cells either respond differently to the murine microenvironment or they provide their own cytokines in a paracrine fashion. Differentiation into erythroid, myeloid and megakaryocytic lineages was also demonstrated by isoelectric focusing (IEF) gel electrophoresis and in situ hybridization for human specific Alu-1 sequences. The engraftment was quantitated by the in situ hybridization technique and the human specific CD45 flow cytometry. Quantitatively significant engraftment could be maintained for long terms (up to week thirty) in the injected animals providing evidence for the multipotency of the HUCB derived highly immature cells to reconstitute hematopoiesis in the SCID mice. In contrast to the previous and concurrent studies where only unseparated or mononuclear cell populations were shown to engraft in the SCID mice, this study provides data on multilineage engraftment of the most immature HUCB derived lineage negative and Rhodamine123-dull cell populations. Besides the long term engraftment as proof for a self renewing cell population in the isolates, serial transplantation of cells from one engrafted SCID mouse to another was also carried out up to the third generation of mice. In the serial transplantation group, positive human engraftment was observed at week 32 in the SCID mice. In a small number of animals, cells infected with a recombinant adeno-associated viral (rAAV) vector containing Fanconi anemia group-C (FACC) gene linked to a neomycinResistant (Neo gene were injected. Using NeoR-specific primers, PCR analysis of the genomic DNA from organs of the SCID mice was performed to assess engraftment of the infected cells. In this group, about half of the animals showed the presence of the neomycin resistant gene. Ultrastructural analysis of the stem cell fractions was also performed but no conclusive results could be obtained. The data from these experiments suggest that the enriched fractions (both non-manipulated and genetically manipulated) from HUCB contain HSCs with properties of engraftment, differentiation and self renewal. This in vivo data on hematopoiesis by the HUCB derived highly immature stem cell fractions will be useful to gain new insights into the biology of hematopoiesis and to evaluate the gene transfer potential of the cells for a variety of clinical disorders.

موضوع مستند نشده

Biological sciences

موضوع مستند نشده

Cellular biology

موضوع مستند نشده

Cellular biology

موضوع مستند نشده

Molecular biology

مستند نام اشخاص تاييد نشده

N.-U.-I. Hashmi

مستند نام اشخاص تاييد نشده

R. E. Morris

نام الکترونيکي

فرمت انتشار

p

نوع ماده

[Thesis]

کد کاربرگه

276903

سطح دسترسي

a

تكميل شده

Y